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学科主题: 物理化学
题名: SECRETORY EXPRESSION OF THE EXO-INULINASE FROM K. MARXIANUS CBS 6556 IN PICHIA PASTORIS
作者: Zhang SF(张素芳) ;  Yang F(杨帆) ;  Wang Q(王倩) ;  Zhao ZB(赵宗保)
会议名称: 13th International Biotechnology Symposium and Exhibition
会议日期: 2008-10-12
出版日期: 2008-10-12
会议地点: 中国
通讯作者: 赵宗保
ISSN: 0168-1656
部门归属: 十八室
主办者: 中国科学院,中国工程院,中国教育部,中国科技部
摘要: Inulin is linear oligosaccharides [1]. Although inulin can be hydrolyzed to monosaccharides non-enzymatically by acid treatment, inhibitory by-products produced that complicate downstream biological process. Therefore, it is pivotal to establish a cost-effective inulinase (EC 3.2.1.7) production. In this study, we isolated the DNA sequence encoding the mature peptide sequence of the exo-inulinase gene from Kluyveromyces marxianus CBS 6556 and expressed in Pichia pastoris X-33. The purified recombinant enzyme (reINU) gave a specific activity of 13100 U•mg−1, which was about 100-fold higher that reported for the wild type protein isolated from K. marxianus CBS6556 [2, 3]. Ferguson plot analysis showed that secretion expressed reINU was a 169 kDa dimer. Detailed analytical assays showed that the optimal temperature and pH for reINU were 60 oC and pH 4.0, respectively. It was found that hydrolysis activity was about 20% higher in the presence of Mn2+. reINU was stable for over 3 days at room temperature. When employed for hydrolysis of the juice of the fresh tuber (water/tuber, 1:1) of Helianthus tuberosus at 50 oC for 1 h, total reduced sugar reached 65 g•l−1, and 85% of the total sugar of the raw material was released based on ion chromatography analysis. In conclusion, we have developed an overproduction system for inulinase that can now be used as biocatalysts for fructose syrups industry and biomass conversion research.
语种: 中文
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内容类型: 会议论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/113172
Appears in Collections:中国科学院大连化学物理研究所_会议论文

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Recommended Citation:
Zhang SF,Yang F,Wang Q,et al. SECRETORY EXPRESSION OF THE EXO-INULINASE FROM K. MARXIANUS CBS 6556 IN PICHIA PASTORIS[C]. 见:13th International Biotechnology Symposium and Exhibition. 中国. 2008-10-12.
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